The effect of culture media and mycotoxin on porcine preimplantation embryonic development has been investigated. In the first experiment, porcine embryonic cleavage rates were similar in NCSU-23 and PZM-3 culture media, while more blastocysts were produced in PZM-3 (P<0.05). BAX and BCL2L1 transcription levels were similar in blastocysts cultured in both media. Cleavage rates were significantly decreased in the presence of cycloheximide (P<0.05), and both á-amanitin and cycloheximide completely inhibited blastocyst formation. In the mycotoxin experiment, porcine embryonic cleavage rates decreased in 10 ìM á-ZEA group, while blastocyst rates decreased in 30 ìM á-ZEA group (P<0.05). Total cell numbers of blastocysts were significantly lower in the 10 µM á-ZEA group (P<0.05). The transcription levels of POU5F1 and BCL2L1 were similar, while that of BAX and the ratio of BAX/BCL2L1 significantly increased in 3 ìM and 10 ìM á-ZEA groups compared to control group.

Zearalenone (ZEN) is an estrogenic mycotoxin produced by fungi in the genus Fusarium. Fusarium species commonly occur in cereal crops such as corn, wheat, barley, and rice which are used in swine feed. In addition to cereals, co-products of ethanol distillation, brewing, and baking are frequently re-purposed as swine feed. ZEN contamination has been related to severe reproductive issues including pelvic organ prolapse (POP), which describes the displacement of pelvic floor organs from their healthy anatomical positions. A recent rise in the rate of POP in sows highlights a need for more information surrounding the occurrence of ZEN in swine feed and the possible role of ZEN in POP. This research aims to understand the relationship between ZEN contamination in swine feed and swine reproductive health. The primary objectives of this work are to (1) characterize the stress relaxation behavior of vaginal tissue in the swine model, (2) determine how ZEN consumption effects the elasticity of pelvic support tissue. (3) determine where ZEN and a related metabolite accumulate in the swine reproductive tract, (4) evaluate the prevalence of ZEN and related Fusarium mycotoxins in swine feed and ingredients, and (5) track ZEN and related Fusarium mycotoxins throughout production of beer and brewers' spent grains (BSG). Results from this work can help stakeholders to mitigate mycotoxin contamination in swine feed, evaluate swine for ZEN toxicosis, and provide care for animals experiencing POP. / Doctor of Philosophy / Under certain conditions, some fungi produce harmful toxins (mycotoxins) that can compromise the health of humans and animals. Mycotoxin-producing fungi are common in cereal crops such as corn, wheat, and barley which may be fed to humans or animals. Swine are especially sensitive to mycotoxins, making feed composition important for animal welfare and profitability. Zearalenone (ZEN) is a mycotoxin that behaves like estrogen, a hormone frequently associated with female reproductive health. Pelvic organ prolapse (POP) is a condition, common in women, wherein organs in the pelvic floor such as the vagina, rectum, and uterus, descend from their healthy anatomical position. Swine herds are experiencing higher rates of POP, which may be related to ZEN contamination in feed. The overall goal of this research is to understand the relationship between mycotoxin contamination in feed and swine reproductive health. The specific objectives of this research are to (1) characterize the mechanical properties of vaginal tissue in swine, (2) characterize mechanical properties of pelvic support tissue associated with POP, in swine fed ZEN, (3) determine which reproductive tissues which accumulate ZEN in swine, (4) evaluate the prevalence of ZEN and related mycotoxins in swine feed, and (5) track ZEN and related mycotoxins throughout production of beer and brewers' spent grain, a co-product used in animal feed. This research can help animal producers develop strategies to mitigate mycotoxin contamination in feed and improve veterinary care.

Doctor of Philosophy / Department of Animal Sciences and Industry / Duane L. Davis / Jim L. Nelssen / A total of 188 sows and their litters were used in 2 experiments to evaluate methods to induce estrus and ovulation in lactating sows and effects on pig growth. In Exp. 1, an altered suckling method (ALT) was designed to combine split-weaning and intermittent suckling as a means to reduce the suckling stimulus in primi- and multiparous sows during the last week of lactation (d 18 to 25). The ALT sows were also removed for daily boar exposure. The ALT treatment produced lactational estrus in 75% and 95% of primiparous and multiparous sows, respectively. The ALT sows were in estrus earlier (P < 0.01) than controls post-farrowing, with no effect on subsequent reproductive performance. From d 18 to 32, the ALT treatment benefited (P < 0.01) growth of lightweight pigs but decreased (P < 0.01) BW gain of heavyweight pigs, resulting in overall similar growth. However, variation in BW was reduced (P < 0.01) by 50% for ALT litters. In Exp. 2, varying suckling reduction strategies were applied to boar-exposed lactating sows. Overall, 76% of sows in suckling reduction treatments expressed estrus in lactation. Split-weaned and ALT sows performed reproductively similar to controls, whereas sows with daily litter separation or a single 24 h litter removal tended (P < 0.10) to have reduced conception rates versus controls or split-weaned sows. Reduced suckling treatments differed in their ability to induce lactational estrus and impact on pig BW gain immediately post-weaning. However, no evidence was found of benefit for pig growth to market weight or litter BW variation. Four additional experiments using 902 nursery pigs were conducted to test the efficacy of potential detoxifying agents against deoxynivalenol (DON) in swine diets. The effects of DON were not offset by adding an algae-modified montmorillonite clay nor by a proprietary blend of preservatives and clays. However, hydrothermally treating DON-contaminated diets with sodium metabisulfite modified the structure of DON to a non-toxic DON-sulfonate adduct and restored nursery pig growth via improved (P < 0.05) ADG, ADFI and G:F.

Aflatoxins, toxic secondary metabolites produced by some Aspergillus species, are a universal agricultural economic problem and a critical health issue. Despite decades of control efforts, aflatoxin contamination is responsible for a global loss of millions of tons of crops each year. We show that host-induced gene silencing is an effective method for eliminating this toxin in transgenic maize. We transformed maize plants with a kernel-specific RNA interference (RNAi) gene cassette targeting the aflC gene, which encodes an enzyme in the Aspergillus aflatoxin biosynthetic pathway. After pathogen infection, aflatoxin could not be detected in kernels from these RNAi transgenic maize plants, while toxin loads reached thousands of parts per billion in nontransgenic control kernels. A comparison of transcripts in developing aflatoxin-free transgenic kernels with those from nontransgenic kernels showed no significant differences between these two groups. These results demonstrate that small interfering RNA molecules can be used to silence aflatoxin biosynthesis in maize, providing an attractive and precise engineering strategy that could also be extended to other crops to improve food security.

No description available.

Internal fruit rot, caused by Fusarium lactis, is as an important disease of greenhouse sweet pepper. Fungal growth was studied microscopically during anthesis and fruit development. Hyphae were observed on the stigmatal surface one day after inoculation (DAI), and in the transmitting tissues of the style and inside the ovary at 5 and 6 DAI. Symptomless seeds from infected fruits yielded colonies of F. lactis when cultured axenically, and typical disease symptoms were observed when fruits were dissected at 45 DAI. Isolates of F. lactis and the related species F. proliferatum and F. verticillioides, which are also associated with internal fruit rot, produced the mycotoxins beauvericin, moniliformin and fumonisin B1 in various combinations, both in infected fruits and in vitro. These findings suggest that internal fruit rot is initiated through infection of the stigma and style during anthesis, and that mycotoxin contamination of infected fruit could pose a health concern. / Plant Science

著作権情報を一部追加（2023-06-30） / 京都大学 / 新制・論文博士 / 博士(農学) / 乙第13508号 / 論農博第2906号 / 新制||農||1095(附属図書館) / 学位論文||R4||N5408(農学部図書室) / (主査)教授 谷 史人, 教授 入江 一浩, 教授 井上 和生 / 学位規則第4条第2項該当 / Doctor of Agricultural Science / Kyoto University / DFAM

Heat stress is detrimental across many, if not most aspects of animal agriculture. Heat stress can have direct effects on the animal, effects on their environment and effects on the feedstuffs they consume. The work presented here will address direct effects on the animal (using dairy cattle) and effects on their feed (using pigs). Dairy cattle were chosen to study direct effects on the animal because they exhibit unique metabolic adaptions during heat stress. One adaptation of interest is a heat-stress induced state of hyperinsulinemia and hypoglycemia while self-limiting feed intake. In regards to effects on feedstuffs, high ambient temperatures are associated with elevated concentrations of mycotoxins in cereal grains. Mycotoxins are problematic because, at sufficient concentrations, they induce specific, toxic effects. Zearalenone is one such mycotoxin that is particularly problematic for swine, as it is a potent phytoestrogen. As such, it can reduce productivity by disrupting reproductive processes. Although these topics are disparate, the overarching goal of this work is to improve agricultural animal productivity during periods of heat stress. Overall, the three objectives of this thesis were to: 1) isolate the production-related effects of hyperinsulinemia with hypoglycemia from heat stress, 2) determine whether glucose supplementation during heat stress could improve or rescue milk production during heat stress, and finally, 3) investigate reproductive tract morphology following different durations of zearalenone consumption in peri-pubertal gilts. The first experiment compared the effects of thermoneutral, thermoneutral + hyperinsulinemic-hypoglycemic clamp (HHC), environmental heat stress, and heat stress + euglycemic clamp (EC) on milk production, milk components, and circulating glucose concentrations. Milk production, as expected, did decrease under heat stress when compared to thermoneutral conditions. Milk production during the HHC was intermediate, which was likely the result of the induced hypoglycemia. Interestingly, the glucose infused during the EC did not improve milk production over heat stress. Baseline blood glucose concentrations declined during heat stress, and those concentrations measured during the thermoneutral period were correlated with the change in blood glucose (from thermoneutral to heat stress). In the second experiment, zearalenone (6 mg) was fed to peri-pubertal gilts for 0, 7 or 21 days. Gilts were monitored daily for observable signs of zearalenone toxicity, and tissues were harvested after 21 days of treatment. No differences in the lengths or weights of any portions of the reproductive tract were observed. Likewise, there were no differences in body weight, carcass weight, dressing percent or liver weight. In summary, heat stress has varied effects on livestock production and the ultimate outcomes are not always predictable. Future work will be aimed at defining the factors that contribute to this variability. / Master of Science / Heat stress can have direct and indirect consequences that affect the efficiency of livestock production. The overall objective of this work is to improve agricultural animal productivity during periods of heat stress. The first study compared milk production, milk characteristics and blood glucose concentrations in the presence or absence of heat stress. Aspects of productivity were also measured when insulin and/or glucose were administered in a manner that mimicked either heat stress or thermoneutral conditions. Heat stress did cause a decline in circulating blood glucose, feed intake, and milk production. Infusion of insulin during thermoneutral conditions caused a decline in milk production but not in feed intake. As a way to compensate for the naturally low circulating glucose during heat stress, glucose was infused during heat stress in the last portion of the experiment. Simply infusing glucose to return the animal to pre-heat stress concentrations did not improve milk production. Further work is needed to determine factors (other than feed intake) that limit milk production during heat stress. The second study investigated whether the mycotoxin, zearalenone, affects the size or weight of the pig's reproductive tract. If differences existed, they would be indicative of potential problems with the function of those tissues. Young female pigs consumed zearalenone for 0, 7 or 21 days and tissues were evaluated 21 days after the experiment began. No changes in tissue weights or lengths were observed. Future experiments should be aimed at determining the concentration and duration of exposure at which zearalenone becomes problematic for young, female pigs.

No description available.

O objetivo da pesquisa foi avaliar os efeitos da AFB1 e FB1, isoladas e associadas e com ou sem a inclusão de um antioxidante (cúrcuma) em frangos de corte, utilizando-se os seguintes tratamentos (T): T1 (controle): 0 AFB1 + 0 FB1 + 0 Cúrcuma; T2: 0 AFB1 + 0 FB1 + 222 mg/kg Cúrcuma; T3: 0 AFB1 + 20 mg/kg FB1 + 0 Cúrcuma; T4: 0 AFB1 + 20 mg/kg FB1 + 222 mg/kg Cúrcuma; T5: 0,5 mg/kg AFB1 + 0 FB1 + 0 Cúrcuma; T6: 0,5 mg/kg AFB1 + 0 FB1 + 222 mg/kg Cúrcuma; T7: 0,5 mg/kg AFB1 + 20 mg/kg FB1 + 0 Cúrcuma; T8: 0,5 mg/kg AFB1 + 20 mg/kg FB1 + 222 mg/kg Cúrcuma. Os parâmetros avaliados incluíram: conversão alimentar, ganho de peso, consumo alimentar, proteínas séricas, hematologia, enzimas hepáticas, histopatologia de vísceras, resposta vacinal, resíduos de aflatoxinas e fumonisina em vísceras e músculos. Os dados foram analisados como um fatorial 2 x 2 x 2 por análise de variância. Não houve interação significativa para os três fatores analisados no desempenho zootécnico, porém houve diferença significativa nas dietas contendo aflatoxina, quando comparada com as outras dietas, para todas as variáveis analisadas. A AST e LDH foram as únicas variáveis significativas para a interação de todos os fatores (AFB1, FB1 e CMT), porém as outras variáveis, exceto a GGT, apresentaram diferença significativa nas dietas contendo aflatoxina, quando comparada com as outras dietas, para todas as variáveis analisadas. As lesões histopatológicas no fígado, rim e bursa de Fabricius foram aumentando gradualmente do tratamento controle até o tratamento contendo adição de aflatoxina e fumonisina, com ou sem cúrcuma, sendo que o efeito mais severo foi observado nos órgãos desses tratamentos. Não houve diferença significativa com a inclusão da cúrcuma. Não houve nenhuma variável significativa para a interação de todos os fatores avaliados para a biometria dos órgãos, porém fígado, rim e coração apresentaram diferença significativa nas dietas contendo aflatoxina, quando comparada com as outras dietas, para todas as variáveis analisadas. Nenhuma variável foi significativa para a interação de todos os fatores avaliados para o hemograma aos 21 dias, porém aos 42 dias, leucócitos, linfócitos e basófilos apresentaram diferença significativa para a interação de todos os fatores. Não houve diferença estatística para a interação dos três fatores analisados para os títulos de anticorpos para a doença de Newcastle. Os resíduos de FB2 no fígado e AFB2 no músculo peitoral foram as únicas variáveis significativas para a interação de todos os fatores analisados. Os resultados obtidos indicam que não houve efeito da cúrcuma na diminuição dos efeitos deletérios da aflatoxina e fumonisina. / The objective of this research project was to evaluate the effects of AFB1 and FB1, alone or in combination and associated or not with an antioxidant (turmeric) in broiler chickens, using the following treatments (T): T1 (control): 0 AFB1 + 0 FB1 + 0 Turmeric; T2: 0 AFB1 + 0 FB1 + 222 mg/kg Turmeric; T3: 0 AFB1 + 20 mg/kg FB1 + 0 Turmeric; T4: 0 AFB1 + 20 mg/kg FB1 + 222 mg/kg Turmeric; T5: 0.5 mg/kg AFB1 + 0 FB1 + 0 Turmeric; T6: 0.5 mg/kg AFB1 + 0 FB1 + 222 mg/kg Turmeric; T7: 0.5 mg/kg AFB1 + 20 mg/kg FB1 + 0 Turmeric; T8: 0.5 mg/kg AFB1 + 20 mg/kg FB1 + 222 mg/kg Turmeric. The parameters evaluated included: feed conversion, weight gain, food intake, serum proteins, hematology, liver enzymes, histopathology of viscera, vaccine response and residues of aflatoxin and fumonisin in tissues of viscera and muscles. Data were analyzed as a 2 x 2 x 2 factorial by analysis of variance. There was no significant interaction for the three analyzed factors on the performance, but there were significant differences in the diets containing aflatoxin when compared to the other diets, for all variables analyzed. AST and LDH were the only significant variables for the interaction of all factors (AFB1, FB1 e CMT), but other variables, with the exception of GGT, showed significant difference in diets containing aflatoxin when compared to other diets for all the variables analyzed. Histopathological lesions in the liver, kidney and bursa of Fabricius were gradually increased from the control treatment to aflatoxin and fumonisin treatment, with or without turmeric, with the most severe effect being observed in the organs of these treatments. There was no significant variable for the interaction of all factors evaluated for organ biometry, but liver, kidney and heart showed a significant difference in diets containing aflatoxin when compared to other diets for all variables analyzed. No variable was significant for the interaction of all factors evaluated for the blood test at 21 days, but at 42 days leukocytes, lymphocytes and basophils showed significant differences for the interaction of all factors. There was no statistical difference for the interaction of all factors analyzed for antibody titers to Newcastle disease. The residues of FB2 in the liver and AFB2 in the pectoral muscle were significant for the interaction of all the factors. The results showed no effect of turmeric powder in reducing the deleterious effects of aflatoxin and fumonisin.